A protocol for simultaneous Ca²⁺ and morphology imaging of brain endothelial tip cells in larval zebrafish

Endothelial tip cells (ETCs) located at growing blood vessels display high morphological dynamics and associated intracellular Ca²⁺ activities with different spatiotemporal patterns during migration. Examining the Ca²⁺ activity and morphological dynamics of ETCs will provide an insight for understanding the mechanism of vascular development in organs, including the brain. Here, we describe a method for simultaneous monitoring and relevant analysis of the Ca²⁺ activity and morphology of growing brain ETCs in larval zebrafish. For complete details on the use and execution of this protocol, please refer to Liu et al. (2020).